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Antifreeze glycopeptide analogues: microwave-enhanced synthesis and functional studies

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Abstract

Antifreeze glycoproteins enable life at temperatures below the freezing point of physiological solutions. They usually consist of the repetitive tripeptide unit (-Ala-Ala-Thr-) with the disaccharide α-d-galactosyl-(1–3)-β-N-acetyl-d-galactosamine attached to each hydroxyl group of threonine. Monoglycosylated analogues have been synthesized from the corresponding monoglycosylated threonine building block by microwave-assisted solid phase peptide synthesis. This method allows the preparation of analogues containing sequence variations which are not accessible by other synthetic methods. As antifreeze glycoproteins consist of numerous isoforms they are difficult to obtain in pure form from natural sources. The synthetic peptides have been structurally analyzed by CD and NMR spectroscopy in proton exchange experiments revealing a structure as flexible as reported for the native peptides. Microphysical recrystallization tests show an ice structuring influence and ice growth inhibition depending on the concentration, chain length and sequence of the peptides.

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Abbreviations

AFGP:

Antifreeze glycopeptide

AFP:

Antifreeze protein

br:

Broad

CD:

Circular dichroism

DIPEA:

N,N-Diisopropylethylamine

DMF:

N,N-Dimethylformamide

EtOAc:

Ethyl acetate

Fmoc:

N-(9H-Fluoren-9-yl)-methoxycarbonyl

GalNAc:

2-Acetamido-2-deoxy-d-galactopyranosyl-

HATU:

O-(7-Azabenzotriazol-1-yl)-N,N,N’,N’-tetramethyluronium hexafluorophosphate

HOAt:

1-Hydroxy-7-azabenzotriazole

MeOH:

Methanol

HOBt:

1-Hydroxybenzotriazole

NaOMe:

Sodium methoxide

NMP:

1-Methyl-2-pyrrolidone

TBTU:

O-(Benzotriazol-1-yl)-N,N,N’,N’-tetramethyluronium tetrafluoroborate

TFA:

Trifluoroacetic acid

TFE:

2,2,2-Trifluoroethanol

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Acknowledgments

The authors gratefully acknowledge support from DFG (SFB 613).

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Authors and Affiliations

  1. Organic and Bioorganic Chemistry, Bielefeld University, Universitätsstr. 25, 33615, Bielefeld, Germany

    Carolin Heggemann, Benjamin Schomburg, Marco Wißbrock & Norbert Sewald

  2. Physical Chemistry, Bielefeld University, Universitätsstr. 25, 33615, Bielefeld, Germany

    Carsten Budke & Thomas Koop

  3. Institute of Chemistry, Eötvös University, 1117, Budapest, Hungary

    Zsuzsa Majer

Authors
  1. Carolin Heggemann
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  2. Carsten Budke
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  3. Benjamin Schomburg
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  4. Zsuzsa Majer
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  5. Marco Wißbrock
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  6. Thomas Koop
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  7. Norbert Sewald
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Correspondence to Norbert Sewald.

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Heggemann, C., Budke, C., Schomburg, B. et al. Antifreeze glycopeptide analogues: microwave-enhanced synthesis and functional studies. Amino Acids 38, 213–222 (2010). https://doi.org/10.1007/s00726-008-0229-0

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  • DOI: https://doi.org/10.1007/s00726-008-0229-0

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